Single-Site Template Nanoparticles for Monoclonal SBS Clustering

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Solution Overview

Problem

Current sequencing by synthesis (SBS) technologies face inefficiencies due to suboptimal use of substrate surface area for seeding and clustering of template polynucleotides, leading to increased time, cost, and complexity in data processing, particularly when clusters are not spatially distinct or polyclonal.

Innovation Solution

The use of nanoparticles with a single template site for bonding a template polynucleotide and multiple accessory sites for accessory oligonucleotides, promoting monoclonal clustering by ensuring each template polynucleotide is attached to a separate nanoparticle, thereby reducing polyclonal clusters and maximizing substrate surface utilization.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If template polynucleotides are seeded densely on substrate surface to maximize surface area utilization, then sequencing throughput increases, but polyclonal clusters form reducing sequencing accuracy

Engineering Contradiction:
Improvesequencing throughputVSAvoidsequencing accuracy
Core Design Contradiction:
ProductivityVSMeasurement precision

Solution Approach 1:

The patent introduces nanoparticles as intermediary carriers that bind template polynucleotides before they are deposited on the substrate. Each nanoparticle acts as a separate seeding unit with a single template polynucleotide, ensuring that even at high densities, each cluster remains monoclonal. This intermediary approach allows dense seeding while maintaining sequencing accuracy by preventing template polynucleotide proximity issues.

Inventive Principle:
Principle #24Intermediary (Mediator)

2Area of stationary object

If multiple template polynucleotides seed close together on substrate, then substrate surface area is utilized efficiently, but spatially distinct clusters cannot be resolved

Engineering Contradiction:
Improvesubstrate surface area utilizationVSAvoidcluster spatial resolution
Core Design Contradiction:
Area of stationary objectVSDifficulty of detecting and measuring

Solution Approach 1:

The patent segments the template polynucleotide binding function across multiple nanoparticle carriers rather than direct substrate binding. Each nanoparticle is spaced apart on the substrate, creating physically separated seeding sites. This segmentation ensures that even though nanoparticles are distributed densely to maximize surface area, the actual template polynucleotide clusters remain spatially distinct and resolvable by imaging systems.

Inventive Principle:
Principle #1Segmentation

3Quantity of substance

If conventional seeding methods are used to maximize template polynucleotide attachment, then sequencing information quantity increases, but data processing complexity increases

Engineering Contradiction:
Improvesequencing information quantityVSAvoiddata processing complexity
Core Design Contradiction:
Quantity of substanceVSDevice complexity

Solution Approach 1:

By using nanoparticles as intermediaries, the patent creates a one-to-one correspondence between nanoparticles and template polynucleotides, simplifying the relationship between physical features and sequence data. Each nanoparticle's position and identity can be tracked through the sequencing process, creating straightforward linkage between spatial coordinates and sequence information, thereby reducing data processing complexity while maintaining high sequencing information quantity.

Inventive Principle:
Principle #24Intermediary (Mediator)

Applied Scientific Principles

This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.

Function Achieved in This Case

This approach enhances the efficiency of SBS by increasing the seeding density and monoclonality of clusters, reducing time and cost, and simplifying data analysis by ensuring clear separation of clusters, thus improving sequencing outcomes.

Implementation Method 1

a single template site for bonding a template polynucleotide to the scaffold selected from a covalent template bonding site and a noncovalent template bonding site

Methodology Applied
Scientific EffectCovalent bonding: Chemical Bonding

Implementation Method 2

a single template site for bonding a template polynucleotide to the scaffold selected from a covalent template bonding site and a noncovalent template bonding site

Methodology Applied
Scientific EffectNoncovalent bonding: Van der Waals Force

Implementation Method 3

a plurality of accessory sites for bonding accessory oligonucleotides to the scaffold selected from covalent accessory oligonucleotide bonding sites and noncovalent accessory oligonucleotide bonding sites

Methodology Applied
Scientific EffectCovalent bonding: Chemical Bonding

Implementation Method 4

a plurality of accessory sites for bonding accessory oligonucleotides to the scaffold selected from covalent accessory oligonucleotide bonding sites and noncovalent accessory oligonucleotide bonding sites

Methodology Applied
Scientific EffectNoncovalent bonding: Van der Waals Force

Data Source

PatentEP4081657B1Nanoparticle with single site for template polynucleotide attachment
Publication Date: 2025.10.29 ILLUMINA INC
  • EP4081657B1 patent drawingFigure 1
  • EP4081657B1 patent drawingFigure 2
  • EP4081657B1 patent drawingFigure 3

AI summary

Provided is a nanoparticle including a scaffold, a single template site for bonding a template polynucleotide to the scaffold, and a plurality of accessory sites for bonding accessory oligonucleotides to the scaffold, wherein the scaffold is selected from an asymmetrical acrylamide polymer one or a dendrimer including lysyl constitutional repeating units, the single template site for bonding a template polynucleotide to the scaffold is selected from a covalent template bonding site and a noncovalent template bonding site and the plurality of accessory sites for bonding accessory oligonucleotides to the scaffold are selected from covalent accessory oligonucleotide bonding sites and noncovalent accessory oligonucleotide bonding sites. Also provided are methods of using the nanoparticle.